Product nameAnti-alpha Tubulin (acetyl K40) antibody [6-11B-1]
See all alpha Tubulin primary antibodies
DescriptionMouse monoclonal [6-11B-1] to alpha Tubulin (acetyl K40)
Specificityab24610 detects acetylated alpha tubulin.
Tested applicationsSuitable for: Flow Cyt, WB, IHC-P, ICC/IF, IHC-Frmore details
Species reactivityReacts with: Mouse, Rat, Sheep, Human, Monkey, Sea urchin
Predicted to work with: Cow
Tissue, cells or virus corresponding to alpha Tubulin.
EpitopeThe antibody recognizes an epitope located on the a3 isoform of Chlamydomonas axonemal a-tubulin, within four residues of Lys40 when this amino acid is acetylated.
- In Western Blot, this antibody gave a positive signal in mouse brain tissue lysate and in the following whole cell lysates: HeLa; NIH3T3; PC12.
Production of this antibody has been changed on 8th April 2016. This antibody is now purified from tissue culture supernatant. This shouldn’t affect the use of this antibody but if you have any issues, please contact our Scientific Support team.
This antibody binds to primary cilia, centrioles, mitotic spindles, midbodies and to subsets of cytoplasmic microtubules in 3T3 and HeLa cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.097% Sodium azide
Concentration information loading...
Purification notesPurified from Tissue culture supernatant.
Light chain typekappa
- Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (ab176560)
- Anti-alpha Tubulin (acetyl K40) antibody [EPR16772] (ab179484)
- Anti-alpha Tubulin antibody [EPR13799] (ab184970)
- Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (HRP) (ab185067)
- Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Alexa Fluor® 647) (ab190573)
- Anti-alpha Tubulin antibody [EP1332Y] - BSA and Azide free (ab216650)
- Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866)
Our Abpromise guarantee covers the use of ab24610 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µl for 106 cells.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 0.03 - 0.06 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC/IF||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
FunctionTubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain.
Sequence similaritiesBelongs to the tubulin family.
modificationsSome glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
Acetylation of alpha chains at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling.
Cellular localizationCytoplasm > cytoskeleton.
- Information by UniProt
- Alpha-tubulin 1 antibody
- ALS22 antibody
- B ALPHA 1 antibody
All lanes : Anti-alpha Tubulin (acetyl K40) antibody [6-11B-1] (ab24610) at 5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 4 : Brain (Mouse) Tissue Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Additional bands at: 140 kDa, 25 kDa, 35 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 150 seconds
ab24610 staining Acetylated alpha Tubulin in monkey kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 3% PFA + 0.1% GA and blocked with 3% BSA + 0.5% Triton X-100 for 45 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 3% BSA + 0.5% Triton X-100) for 1 hour at 21°C. An Alexa Fluor® 647-conjugated donkey anti-rabbit IgG polyclonal (2 µg/ml) was used as the secondary antibody.
ab24610 at 1/100 dilution staining acetylated alpha tubulinin in prostate carcinoma by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Sections were paraformaldehyde fixed, permeabilized in Triton X-100 prior to blocking in 1% serum for 1 hour at 27°C and then incubated with ab24610 for 12 hours at 4°C. Alexa Fluor® 546 donkey polyclonal to mouse Ig, diluted 1/500, was used as the secondary antibody.
ICC/IF image of ab24610 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab24610, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT™ FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
ICC/IF image of ab24610 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24610, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Overlay histogram showing HeLa cells stained with ab24610 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab24610, 1µg/1x106 cells) for 30 min at 22ºC. (This data was generated from a purified version of the antibody. Some lots are produced as ascites fluid. We suggest 1µl/1x106 cells for ascites preparations). The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This product has been referenced in:
- Corrigan MA et al. TRPV4-mediates oscillatory fluid shear mechanotransduction in mesenchymal stem cells in part via the primary cilium. Sci Rep 8:3824 (2018). Read more (PubMed: 29491434) »
- Yurchenko OV et al. Nervous system development in the Pacific oyster, Crassostrea gigas (Mollusca: Bivalvia). Front Zool 15:10 (2018). Read more (PubMed: 29681988) »